Materials & Methods: Results: Figure 1(a&b): Front and back view of plate with bacteria only (VDS JWS XBG 2/27/14 NONE BL21(DE3)) the evening of Day 2. No bacterial growth evident.
Figure 2(a&b): Front and back view of plate with bacteria and ampicillin-resistant, purple encoding protein plasmid (VDS JWS XBG 2/27/14 AMP BL21(DE3) pGEM-gbr22) the evening of day 2. No bacterial growth evident.
Figure 3: VDS JWS XBG 2/27/14 FUN plate with bacteria from coughing. Photo was taken evening of day 2. No bacterial growth was evident.
Figure 4: "VDS ARW AL 2/27/14 BL21(DE3) SOC media NO DNA" plate with bacteria only. Photo taken evening of day 2. No bacterial growth evident.
Figure 5: "VDS ARW AL 2/27/14 BL21(DE3) pGEM-gbr22 soc media" plate with bacteria and ampicillin-resistant, purple encoding protein plasmid inserted in bacteria. Photo taken evening of day 2. Bacterial growth was evident with many single colonies present.
Figure 6 (a&b): Flasks containing LB, ampicillin, BL21(DE3)+pGEM-gbr22 (VDS xbg 2/28/14), covered with foil after spending 24hrs in shaking incubator (37 degrees Celsius & 200-350 rpm). Photo taken 6:10pm on day 3. 5a is one flask with yellow media, indicating no bacterial growth. 5b is the other flask with purple media, indicating bacterial growth.
Figure 7: Wet pellet of bacteria (BL21(DE3)) with purple protein (encoded by plasmid pGEM-gbr22). Photo taken day 3 after centrifuge and disposing of extra liquid media.
Figure 8: "Sample 5" indicates the Elution 1 tube, containing the majority of the purified gbr22 protein (evident by purplish color). "Sample 6" indicates the Elution 2 tube, containing some purified gbr22 protein (evident by more clear color).
Figure 9. Trial 1 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 280nm, using Protein - A280 mode. The absorbance shows to be 0.237 A280.
Figure 10. Trial 2 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 280nm, using Protein - A280 mode. The absorbance shows to be 0.237 A280.
Figure 11. Trial 1 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 574nm as well as at 280nm, using UV/VIS mode. Because this mode uses a 1mm path length instead of 1cm, the absorbance reading at 574 nm, multiplied by 10, is 0.18 A574 (while for 280nm, it is 0.33 A574).
Figure 12. Trial 2 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 574nm as well as at 280nm, using UV/VIS mode. Because this mode uses a 1mm path length instead of 1cm, the absorbance reading at 574 nm, multiplied by 10, is 0.14 A574 (while for 280nm, it is 0.31 A574).
Figure 13. Gel of molecular weight standards (1st column; ColorPlus Prestained Lot#0131303, New England Biolabs), another student's samples 1-6 (columns 2-7), and my samples 4-6 (columns 8-10), before drying.
Figure 14. Gel of molecular weight standards (1st column; ColorPlus Prestained Lot#0131303, New England Biolabs), another student's samples 1-6 (columns 2-7), and my samples 4-6 (columns 8-10), after drying. Both sample sets were labeled based on what was contained: cell lysate in column 2, soluble fraction in column 3, flow through in column 4, wash in column 5, elution 1 in column 6, elution 2 in column 7, wash in column 8, elution 1 in column 9, and elution 2 in column 10. The molecular weight standards measurements/indicators (ColorPlus Prestained Log #0131303, New England Biolabs) are shown to the left of the gel.
Introduction:
Materials & Methods:
Results:
Figure 1(a&b): Front and back view of plate with bacteria only (VDS JWS XBG 2/27/14 NONE BL21(DE3)) the evening of Day 2. No bacterial growth evident.
Figure 2(a&b): Front and back view of plate with bacteria and ampicillin-resistant, purple encoding protein plasmid (VDS JWS XBG 2/27/14 AMP BL21(DE3) pGEM-gbr22) the evening of day 2. No bacterial growth evident.
Figure 3: VDS JWS XBG 2/27/14 FUN plate with bacteria from coughing. Photo was taken evening of day 2. No bacterial growth was evident.
Figure 4: "VDS ARW AL 2/27/14 BL21(DE3) SOC media NO DNA" plate with bacteria only. Photo taken evening of day 2. No bacterial growth evident.
Figure 5: "VDS ARW AL 2/27/14 BL21(DE3) pGEM-gbr22 soc media" plate with bacteria and ampicillin-resistant, purple encoding protein plasmid inserted in bacteria. Photo taken evening of day 2. Bacterial growth was evident with many single colonies present.
Figure 6 (a&b): Flasks containing LB, ampicillin, BL21(DE3)+pGEM-gbr22 (VDS xbg 2/28/14), covered with foil after spending 24hrs in shaking incubator (37 degrees Celsius & 200-350 rpm). Photo taken 6:10pm on day 3. 5a is one flask with yellow media, indicating no bacterial growth. 5b is the other flask with purple media, indicating bacterial growth.
Figure 7: Wet pellet of bacteria (BL21(DE3)) with purple protein (encoded by plasmid pGEM-gbr22). Photo taken day 3 after centrifuge and disposing of extra liquid media.
Figure 8: "Sample 5" indicates the Elution 1 tube, containing the majority of the purified gbr22 protein (evident by purplish color). "Sample 6" indicates the Elution 2 tube, containing some purified gbr22 protein (evident by more clear color).
Figure 9. Trial 1 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 280nm, using Protein - A280 mode. The absorbance shows to be 0.237 A280.
Figure 10. Trial 2 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 280nm, using Protein - A280 mode. The absorbance shows to be 0.237 A280.
Figure 11. Trial 1 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 574nm as well as at 280nm, using UV/VIS mode. Because this mode uses a 1mm path length instead of 1cm, the absorbance reading at 574 nm, multiplied by 10, is 0.18 A574 (while for 280nm, it is 0.33 A574).
Figure 12. Trial 2 of absorbance reading of Elution 1 solution by nanodrop spectrophotometer at 574nm as well as at 280nm, using UV/VIS mode. Because this mode uses a 1mm path length instead of 1cm, the absorbance reading at 574 nm, multiplied by 10, is 0.14 A574 (while for 280nm, it is 0.31 A574).
Figure 13. Gel of molecular weight standards (1st column; ColorPlus Prestained Lot#0131303, New England Biolabs), another student's samples 1-6 (columns 2-7), and my samples 4-6 (columns 8-10), before drying.
Figure 14. Gel of molecular weight standards (1st column; ColorPlus Prestained Lot#0131303, New England Biolabs), another student's samples 1-6 (columns 2-7), and my samples 4-6 (columns 8-10), after drying. Both sample sets were labeled based on what was contained: cell lysate in column 2, soluble fraction in column 3, flow through in column 4, wash in column 5, elution 1 in column 6, elution 2 in column 7, wash in column 8, elution 1 in column 9, and elution 2 in column 10. The molecular weight standards measurements/indicators (ColorPlus Prestained Log #0131303, New England Biolabs) are shown to the left of the gel.
Discussion:
Conclusions:
References: